• Title/Summary/Keyword: HPLC analysis

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Isolation and Identification of Adventitious Root Formation Inducing Substances from Cortex of cinnamomum cassia J.Presl (육계(Cortex of Cinnamomum cassia J.Presl) 추출물로부터 부정근 형성 유도물질 분리 및 동정)

  • Joo Ho Yeo;Jeong Kyu Baek;Jee Sung Park;Kun Woo Kim
    • Korean Journal of Plant Resources
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    • v.37 no.1
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    • pp.11-21
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    • 2024
  • In this study, as a result of exploring the physiological activity of plants useful for agriculture on various plant resources, it was possible to confirm an activity similar to auxin that promotes plant rooting in methanol extract of Cinnamon Bark (cortex of Cinnamomum cassia J.Presl). After separating the active body by applying column chromatography and HPLC to the CHCl3 active fraction obtained by solvent extraction for each polarity from the methanol extract of cinnamon bark, cinnamyl alcohol was identified through GC/MS analysis. By bioassay using cinnamyl alcohol standard and the active fraction separated and purified from the methanol extract of cinnamon bark, the rooting rate of mung bean seedlings of the cinnamyl alcohol standard was 290% compared with the untreated control at 134.2 ㎍/mL concentration, and the adventitious root formation activity similar to the rooting rate (268.6%; 100 ㎍/mL) of the active fraction was shown. In conclusion, it is believed that cinnamyl alcohol contained in methanol extract of Cinnamon Bark is the main compound that induces adventitious root formation in mung bean.

Ginseng root-derived exosome-like nanoparticles protect skin from UV irradiation and oxidative stress by suppressing activator protein-1 signaling and limiting the generation of reactive oxygen species

  • Wooram Choi;Jeong Hun Cho;Sang Hee Park;Dong Seon Kim;Hwa Pyoung Lee;Donghyun Kim;Hyun Soo Kim;Ji Hye Kim;Jae Youl Cho
    • Journal of Ginseng Research
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    • v.48 no.2
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    • pp.211-219
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    • 2024
  • Background: Recently, plant-derived exosome-like nanoparticles (PDENs) have been isolated, and active research was focusing on understanding their properties and functions. In this study, the characteristics and molecular properties of ginseng root-derived exosome-like nanoparticles (GrDENs) were examined in terms of skin protection. Methods: HPLC-MS protocols were used to analyze the ginsenoside contents in GrDENs. To investigate the beneficial effect of GrDENs on skin, HaCaT cells were pre-treated with GrDENs (0-2 × 109 particles/mL), and followed by UVB irradiation or H2O2 exposure. In addition, the antioxidant activity of GrDENs was measured using a fluorescence microscope or flow cytometry. Finally, molecular mechanisms were examined with immunoblotting analysis. Results: GrDENs contained detectable levels of ginsenosides (Re, Rg1, Rb1, Rf, Rg2 (S), Gyp17, Rd, C-Mc1, C-O, and F2). In UVB-irradiated HaCaT cells, GrDENs protected cells from death and reduced ROS production. GrDENs downregulated the mRNA expression of proapoptotic genes, including BAX, caspase-1, -3, -6, -7, and -8 and the ratio of cleaved caspase-8, -9, and -3 in a dose-dependent manner. In addition, GrDENs reduced the mRNA levels of aging-related genes (MMP2 and 3), proinflammatory genes (COX-2 and IL-6), and cellular senescence biomarker p21, possibly by suppressing activator protein-1 signaling. Conclusions: This study demonstrates the protective effects of GrDENs against skin damage caused by UV and oxidative stress, providing new insights into beneficial uses of ginseng. In particular, our results suggest GrDENs as a potential active ingredient in cosmeceuticals to promote skin health.

Neuroprotective effect of Aster yomena ethanolic extract in HT-22 and SK-N-MC cells based on antioxidant activity

  • In Young Kim;Jong Min Kim;Hyo Lim Lee;Min Ji Go;Han Su Lee;Ju Hui Kim;Hyun Ji Eo;Chul-Woo Kim;Ho Jin Heo
    • Food Science and Preservation
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    • v.31 no.1
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    • pp.99-111
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    • 2024
  • The antioxidant potentials of ethanolic extracts derived from Aster yomena (A. yomena) were evaluated by assessing their total phenolic and flavonoid contents and radical scavenging activities. Our findings revealed that the 60% ethanolic extract of A. yomena exhibited the most robust antioxidant properties among all extracts tested. Specifically, the IC50 values for the 2,2'-azino-bis (3-ethyl benzothiazoline-6-sulfonic acid) and 1,1-diphenyl-2-picrylhydrazyl radical scavenging activities of the 60% ethanolic extract from A. yomena were determined to be 1,640.30 ㎍/mL and 2,655.10 ㎍/mL, respectively. Moreover, the inhibitory effect on malondialdehyde increased with the 60% ethanolic extract from A. yomena. To assess the neuroprotective effects, we examined the impact of the 60% ethanolic extract from A. yomena against H2O2-induced cytotoxicity in HT-22 (mouse hippocampal neuronal cell line) and SK-N-MC (human neuroblastoma cell line) cells. The results demonstrated a significant improvement in cell viability and reduced intracellular oxidative stress. Furthermore, the major bioactive compounds present in the 60% ethanolic extract from A. yomena were identified as chlorogenic acid and rutin through high-performance liquid chromatography (HPLC) analysis.

Vitamin K variations in conger eel (Conger myriaster) influenced by harvest time and size

  • Junhee Park;Kang-Hee Lee;Rea Mae Cuario Templonuevo;Jinwoo Yang;Jiyeon Chun
    • Food Science and Preservation
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    • v.30 no.6
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    • pp.929-943
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    • 2023
  • This study analyzed the variation in vitamin K content in conger eel (Conger myriaster) caught off Tongyeong, Gyeongsangnam-do, Korea, focusing on the influence of size (large and small) and harvest period (monthly throughout 2021). We applied enzymatic extraction and HPLC-fluorescence methods for the analysis of vitamin K1 (phylloquinone) and K2 (menaquinone). The vitamin K content in conger eel varied significantly with size and harvest season (p<0.05). In large-sized samples, the phylloquinone content peaked in July (0.80±0.09 ㎍/100 g), while the highest menaquinone content was in May (0.79±0.11 ㎍/100 g). Conversely, in small-sized conger eels, the highest phylloquinone was found in December (1.94±0.15 ㎍/100 g), and the peak menaquinone level was in January (0.66±0.02 ㎍/100 g). The fat content was highest in July for large samples and in January for smaller ones. There was a positive correlation between fat and total vitamin K contents in conger eel (r=0.631, 0.667). Method validation and quality control measures ensured data reliability for vitamin K1 and K2 analyses. This study provides reliable information on the size and seasonal variations of vitamin K in conger eels, a staple in the Korean diet. This information is valuable for inclusion in Korea's national food nutrition database and for formulating future national health and nutrition policies.

Methanol-involved heterogeneous transformation of ginsenoside Rb1 to rare ginsenosides using heteropolyacids embedded in mesoporous silica with HPLC-MS investigation

  • Mengya Zhao;Yusheng Xiao;Yanyan Chang;Lu Tian;Yujiang Zhou;Shuying Liu;Huanxi Zhao;Yang Xiu
    • Journal of Ginseng Research
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    • v.48 no.4
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    • pp.366-372
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    • 2024
  • Background: The biological activity and pharmacological effects of rare ginsenosides have been proven to be superior to those of the major ginsenosides, but they are rarely found in ginseng. Methods: Ginsenoside Rb1 was chemically transformed with the involvement of methanol molecules by a synthesized heterogeneous catalyst 12-HPW@MeSi, which was obtained by the immobilization of 12-phosphotungstic acid on a mesoporous silica framework. High-performance liquid chromatography coupled with mass spectrometry was used to identify the transformation products. Results: A total of 18 transformation products were obtained and identified. Methanol was found to be involved in the formation of 8 products formed by the addition of methanol molecules to the C-24 (25), C-20 (21) or C-20 (22) double bonds of the aglycone. The transformation pathways of ginsenoside Rb1 involved deglycosylation, addition, elimination, cycloaddition, and epimerization reactions. These pathways could be elucidated in terms of the stability of the generated carbenium ion. In addition, 12-HPW@MeSi was able to maintain a 60.5% conversion rate of Rb1 after 5 cycles. Conclusion: Tandem and high-resolution mass spectrometry analysis allowed rapid and accurate identification of the transformation products through the characteristic fragment ions and neutral loss. Rare ginsenosides with methoxyl groups grafted at the C-25 and C-20 positions were obtained for the first time by chemical transformation using the composite catalyst 12-HPW@MeSi, which also enabled cyclic heterogeneous transformation and facile centrifugal separation of ginsenosides. This work provides an efficient and recyclable strategy for the preparation of rare ginsenosides with the involvement of organic molecules.

Analysis of Isoflavonoid Contents in Astragalus membranaceus Bunge Cultivated in Different Areas and at Various Ages (황기의 산지별 및 연근별 isoflavonoids의 함량분석)

  • Im, Kyung-Ran;Kim, Mi-Jin;Jung, Teak-Kyu;Yoon, Kyung-Sup
    • KSBB Journal
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    • v.25 no.3
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    • pp.271-276
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    • 2010
  • This study was carried out to obtain the basic information for isoflavonoid contents that can be used to index Astragalus membranaceus B. cultivated in the Republic of Korea and China. Isoflavonoid contents in Astragalus membranaceus B. which were cultivated in various areas (Jecheon, Jeongseon, Yeongju, and Taebaek in Korea, and China) and ages (1-year-old, 3-years-old) were determined. Calycosin and formononetin as major constituents were determinated by HPLC method in Astragalus membranaceus B. The results show that there were no statistically significant differences for the average contents of isoflavonoids among 1-year-old and 3-years-old. However, isoflavonoid contents were significant differences according to the cultivation areas. HPLC analysis showed that the calycosin content of 1-year-old at Jeongseon was the highest level of $0.090{\pm}0.002%$ and that of 1-year-old at Yeongju was the lowest level of $0.010{\pm}0.001%$. The highest level of formononetin content was $0.050{\pm}0.001%$ of 1-year-old at China, while the lowest level was $0.020{\pm}0.001%$ of 1-year-old at Yeongju. These results strongly suggest that contents of isoflavonoid in Astragalus membranaceus B. might be quite different with respect to the cultivation areas.

Production of a hypothetical polyene substance by activating a cryptic fungal PKS-NRPS hybrid gene in Monascus purpureus (홍국Monascus purpureus에서 진균 PKS-NRPS 하이브리드 유전자의 발현 유도를 통한 미지 polyene 화합물의 생성)

  • Suh, Jae-Won;Balakrishnan, Bijinu;Lim, Yoon Ji;Lee, Doh Won;Choi, Jeong Ju;Park, Si-Hyung;Kwon, Hyung-Jin
    • Journal of Applied Biological Chemistry
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    • v.61 no.1
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    • pp.83-91
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    • 2018
  • Advances in bacterial and fungal genome mining uncover a plethora of cryptic secondary metabolite biosynthetic gene clusters. Guided by the genome information, targeted transcriptional derepression could be employed to determine the product of a cryptic gene cluster and to explore its biological role. Monascus spp. are food grade filamentous fungi popular in eastern Asia and several genome data belong to them are now available. We achieved transcription activation of a cryptic fungal polyketide synthase-nonribosomal peptide synthase gene Mpfus1 in Monascus purpureus ${\Delta}MpPKS5$ by inserting Aspergillus gpdA promoter at the upstream of Mpfus1 through double crossover gene replacement. The gene cluster with Mpfus1 show a high similarity to those for the biosynthesis of conjugated polyene derivatives with 2-pyrrolidone ring and the mycotoxin fusarin is the representative member of this group. The ${\Delta}MpPKS5$ is incapable of producing azaphilone pigment, providing an excellent background to identify chromogenic and UV-absorbing compounds. Activation of Mpfus1 resulted in a yellow hue on mycelia and its methanol extract exhibit a maximum absorption at 365 nm. HPLC analysis of the organic extracts indicated the presence of a variety of yellow compounds in the extract. This implies that the product of MpFus1 is metabolically or chemically unstable. LC-MS analysis guided us to predict the MpFus1 product and to propose that the Mpfus1-containing gene cluster encode the biosynthesis of a desmethyl analogue of fusarin. This study showcases the genome mining in Monascus and the possibility to unveil new biological activities embedded in it.

Establishment of Pre-Harvest Residue Limit for Pyrimethanil and Methoxyfenozide during Cultivation of grape (포도(Vitis vinifera L.) 중 Pyrimethanil 및 Methoxyfenozide의 생산단계 잔류허용기준 설정)

  • Kim, Ji Yoon;Woo, Min Ji;Hur, Kyung Jin;Manoharan, Saravanan;Kwon, Chan-Hyeok;Hur, Jang Hyun
    • The Korean Journal of Pesticide Science
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    • v.19 no.2
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    • pp.81-87
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    • 2015
  • The present study was aimed to predict the pre-harvest residue limits (PHRLs) of pyrimethanil (fungicide) and methoxyfenozide (insecticide) in grape, and to estimate their biological half-lives and residual characteristics. The pesticides were sprayed once on grape in two different fields 10 days before harvest. At the end of 0, 1, 2, 3, 5, 7 and 10 days after application, samples were harvested for further analysis. The residual pesticides were extracted with acetonitrile and partitioned with dichloromethane, and the high-performance liquid chromatography with diode array detector (HPLC/DAD) was employed for the residue analysis. The results obtained in the present study show that the limit of detection of both pesticides were found to be $0.01mg\;kg^{-1}$. The recoveries of these pesticides were ranged between 80.6% and 102.5% with coefficient of variation lower than 10%. The biological half-lives of both pesticides were observed in field 1 and field 2 which shows 7.7 and 7.4 days for pyrimethanil and 5.1 and 6.1 days for methoxyfenozide, respectively. Further, the PHRL of pyrimethanil and methoxyfenozide was found to be $8.90mg\;kg^{-1}$ and $5.51mg\;kg^{-1}$, respectively at 10 days before harvest. Consequently, the present study suggests that the residual amounts of both pesticides will be lower than the maximum residue limits (MRLs) when grape is harvested.

Isolation and Chemical Analysis of Potent Anti-Complementary Polysaccharides from Fruiting Bodies of the Fomes fomentarius (말굽버섯 자실체에서 분리한 항보체 활성 다당체의 화학적 분석)

  • Park, Jung-Keun;Park, Kwe-Won;Shin, Kwang-Soon;Lee, Chang-Muk;Seok, Soon-Ja;Kim, Jeong-Bong;Koo, Bon-Sung;Han, Bum-Soo;Yoon, Sang-Hong
    • Microbiology and Biotechnology Letters
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    • v.41 no.2
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    • pp.198-206
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    • 2013
  • The five anti-complementary polysaccharides (MFKF-NP, MFKF-AP1${\alpha}$, ${\beta}$, and MFKF-AP2${\alpha}$, ${\beta}$) were separated from hot water extracts of fruiting bodies of Fomes fomentarius by two subsequent column chromatography using DEAE-sepharose FF and Concanavalin A-sepharose 4B. The order of anti-complementary activity was MFKF-AP1${\beta}$ > MFKF-AP1${\alpha}$ > MFKF-AP2${\alpha}$ > MFKF-AP2${\beta}$ > MFKF-NP > Polysaccharide Krestine (PSK). Especially, MFKF-AP1${\beta}$ among those showed the most excellent anti-complementary activity (70% of ITCH50 value at $20{\mu}g/ml$). The monosaccharide composition analysis by gas chromatography indicates that MFKF-AP1${\alpha}$ and ${\beta}$ are a kind of homoxylan consisted mainly of xylose above 97%. Molecular weight of MFKF-AP1${\beta}$, major anti-complementary polysaccharide, was estimated to be about 12,000 by high performance liquid chromatography (HPLC). After the incubation of the serum with MFKF-AP1${\beta}$ in the presence or absence of $Mg^{++}$ and $Ca^{++}$ ions, its anti-complementary activity was investigated. This result indicated that MFKF-AP1${\beta}$ seems to be activator both on the classical and the alternative pathway of complement activation.

Antioxidative Effect and Component Analysis of Eriobotrya japonica Leaf Extracts (비파엽 추출물의 항산화 효능과 성분분석)

  • Kim, Su-Ji;Park, Jin-O;Park, Soo-Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.38 no.1
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    • pp.57-65
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    • 2012
  • In the present study, the antioxidative properties, inhibitory activity on tyrosinase, and active components of Eriobotrya japonica (E. japonica) leaf extract were investigated. The free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity ($FSC_{50}$) of extract/fraction of E. japonica leaf was in the order 50 % ethanol extract ($22.625{\mu}g/mL$) < ethyl acetate fraction (6.75) < deglycosylated aglycone fraction (5.06). Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of fraction/extracton ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system using the luminol-dependent chemiluminescenceassay were investigated. $OSC_{50}$ of the ethyl acetate fraction, deglycosylated aglycone fraction, and ethanol extract were 0.75, 0.79, and $1.61{\mu}g/mL$, respectively. The cellular protective effects of E. japonica leaf extract on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The protective effects of extract/fraction of E. japonica leaf were increased in a in a concentration dependent manner ($5{\sim}50{\mu}g/mL$). Especially, ${\tau}50$ of ethyl acetate fraction at concentrations of $10{\mu}g/mL$ and $50{\mu}g/mL$ showed the most protective effects at 390.8 min and 1471.5 min. The inhibitory effect ($IC_50$) on tyrosinase of E. japonica leaf extracts was higher than arbutin, known as a skin-whitening agent. The order of inhibitory effects was acetate fraction ($75.25{\mu}g/mL$) < 50 % extract (74.1) < deglycosylated aglycone fraction (43.35). TLC of the ethyl acetate fraction showed 7 bands (EJL 1 - EJL 7). HPLC of the aglycone fraction exhibited 2 peaks, kaempferol and quercetin. The amounts of kaempferol and quercetin were 53.7 and 46.3 %. respectively. Therefore, The amounts of kaempferol and its glucoside were a little bit higher than quercetin and its glucoside in E. japonica leaf extract. Accordingly, these findings suggest that extracts/fractions of E. japonica leaf can function as antioxidants in biological systems, especially skin exposed to UV radiation, and protect cellular membranes against ROS. Thus, the extract/fraction of E. japonica leaf may be used in novel functional cosmetics as antioxidants against skin photoaging.